Biochemical and growth performance of the aquatic macrophyte Azolla filiculoides to sub-chronic exposure to cylindrospermopsin
Physiological and biochemical effects of cylindrospermopsin (CYN), a cyanobacterial toxin that inhibits protein synthesis and released during a harmful cyanobacterial bloom, has been overlooked in plants. Therefore, at the present research, the toxic effects (physiological and biochemical) of a crude extract containing CYN were assessed in the aquatic fern Azolla filiculoides exposed to three concentrations (0.05, 0.5 and 5 μg CYN mL−1). At 5 μg CYN mL−1, fern growth rate has showed a drastic decrease (0.001 g g−1 day−1) corresponding to a 99.8 % inhibition, but at the concentrations of 0.05 and 0.5 μg CYN mL−1 the growth rate was similar to the control plants. Growth rate also indicated a IC50 of 2.9 μg CYN mL−1. Those data point to the presence of other compounds in the crude extract may stimulate the fern growth and/or the fern is tolerant to CYN. Chlorophyll (a and b), carotenoids and protein content as well as the activities of glutathione reductase (GR) and glutathione-S-transferase (GST) has increased at 5 μg CYN mL−1 which may indicate that photosynthesis and protein synthesis are not affected by CYN and the probable activation of defense and detoxifying mechanisms to overcome the effects induced by the presence of CYN. Low uptake of cylindrospermopsin (1.314 μg CYN g−1 FW) and low bioconcentration factor (0.401) point towards to a safe use of A. filiculoides as biofertilizer and as food source, but also indicate that the fern is not suitable for CYN phytoremediation.
KeywordsAzolla filiculoides Cylindrospermopsin Antioxidative enzymes Growth rate Photosynthetic pigments
This research was partially supported by 1) the European Regional Development Fund (ERDF) through the COMPETE (Operational Competitiveness Programme) and national funds through FCT (Foundation for Science and Technology) under the project PEst-C/MAR/LA0015/2013 and 2) Porto University under the project IJUP2011_3. The European Social Funding (FSE) under the Human Potential Operational Program (POPH) of National Strategic Reference Board (QREN) supports the fellowship SFRH/BPD/44459/2008 to Ana L. Pereira. Thanks to Stephan Haefele and Agnes Padre of IRRI for sending A. filiculoides (FI1001).
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Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Beyer D, Surányi G, Vasas G, Roszik J, Erdődi F, Hamvas M-M, Bácsi I, Bátori R, Serfőző Z, Szigeti ZM, Vereb G, Demeter Z, Gonda S, Máthé C (2009) Cylindrospemopsin induces alterations of root histology and microtubule organization in common reed (Phragmites australis) plantlets cultured in vitro. Toxicon 54:440–449. doi: 10.1016/j.toxicon.2009.05.008 CrossRefGoogle Scholar
- Campos A, Araújo P, Pinheiro C, Azevedo J, Osório H, Vasconcelos V (2013) Effects on growth, antioxidant enzyme activity and levels of extracellular proteins in green alga Chlorella vulgaris exposed to crude cyanobacterial extracts and pure microcystin and cylindrospermospin. Ecotoxicol Environ Saf 94:45–53. doi: 10.1016/j.ecoenv.2013.04.019 CrossRefGoogle Scholar
- Carlberg I, Mannervik B (1975) Purification and characterization of the flavoenzyme glutathione reductase from rat liver. J Biol Chem 250:5475–5480Google Scholar
- Carrapiço F (2010) Azolla as a superorganism. Its implication in symbiotic studies. In: Seckbach J and Grube M (eds) Symbioses and stress: Joint ventures in biology, cellular origin, life in extreme habitats and astrobiology. Springer, Amesterdam, pp 225–241Google Scholar
- Carrapiço F, Teixeira G, Diniz MA (2000) Azolla as a biofertiliser in Africa. A challenge for the future. Revista de Ciências Agrárias 23:120–138Google Scholar
- Chiswell RK, Shaw GR, Eaglesham G, Smith MJ, Norris RL, Seawright AA, Moore MR (1999) Stability of cylindrospermopsin, the toxin from the cyanobacterium, Cylindrospermopsis raciborskii: effect of pH, temperature, and sunlight on decomposition. Environ Toxicol 14:155–161. doi: 10.1002/(SICI)1522-7278(199902 CrossRefGoogle Scholar
- Cruz AA, Hiskia A, Kaloudis T, Chernoff N, Hill D, Antoniou MG, He X, Loftin K, O’Shea K, Zhao C, Peleaz M, Han C, Lynch TJ, Dionysiou DD (2013) A review on cylindrospermopsin: the global occurrence, detection, toxicity and degradation of a potent cyanotoxin. Environ Sci 15:1979–2003. doi: 10.1039/c3em00353a Google Scholar
- Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione-S-transferases: the first enzymatic step in mercapturiac acid formation. J Biol Chem 249:7130–7139Google Scholar
- Khatun A, Ali MA, Dingle JG (1999) Comparison of the nutritive value for laying hens of diets containing azolla (Azolla pinnata) based on formulation using digestible protein and digestible amino acid versus total protein and total amino acid. Animal Feed Sci Technol 81:43–56. doi: 10.1016/S0377-8401(99)00071-1 CrossRefGoogle Scholar
- Kinnear SHW, Duivenvoorden LJ, Fabbro LD (2007) Growth and bioaccumulation in Spirodela oligorrhiza following exposure to Cylindrospermopsis raciborskii whole cell extracts. Aust J Ecotoxicol 13:19–31Google Scholar
- Kotai J (1972) Instruction for preparation of modified nutrient solution Z8 for algae. Publication B-11/69, Norwegian Institute for Water Research, OsloGoogle Scholar
- Máthé C, Vasas G, Borbély G, Erdődi F, Beyer D, Kiss A, Surányi G, Gonda S, Jámbrik K, Hamvas M-M (2013) Histological, cytological and biochemical alterations induced by microcystin-LR and cylindrospermopsin in white mustard (Sinapsis alba L.) seedlings. Acta Biol Hung 64:71–85. doi: 10.1556/ABiol.64.2013.1.7 CrossRefGoogle Scholar
- OECD (2002) OECD guidelines for the testing of chemicals. Revised proposal for a new guideline 221. Lemna sp. growth inhibition test. OECD. http://www.oecd.org/chemicalsafety/testing/1948054.pdf. Accessed 13 June 2014
- Prieto A, Campos A, Cameán A, Vasconcelos V (2011) Effects on growth and antioxidative stress status of rice plants (Oryza sativa) exposed to two extracts of toxin-producing cyanobacteria (Aphanizomenon ovalisporum and Microcystis aeruginosa). Ecotoxicol Environ Saf 74:1973–1980. doi: 10.1016/j.ecoenv.2011.06.009 CrossRefGoogle Scholar
- Sánchez-Viveros G, Ferrera-Cerrato R, Alarcón A (2011) Short-term effects of arsenate-induced toxicity on growth, chlorophyll and carotenoid contents, and total content of phenolic compounds of Azolla filiculoides. Water Air Soil Pollut 217:455–462. doi: 10.1007/s11270-010-0600-0 CrossRefGoogle Scholar
- Vasas G, Gáspár A, Surányi G, Batta G, Gyémánt G, Hamvas M-M, Máthé C, Grigorszky I, Molnár E, Bolbély G (2002) Capillary electrophoretic assay and purification of cylindrospermospin, a cyanobacterial toxin from Aphanizomenon ovalisporum, by plant test (blue-green Sinapsis test). Anal Biochem 302:95–103. doi: 10.1006/abio.2001.5525 CrossRefGoogle Scholar
- Zar JH (1999) Biostatistical analysis. Prentice Hall International Inc, New JerseyGoogle Scholar